Resuspension, Purification, and Preparation of Working Aliquots of Oligos v1

This protocol details the steps for resuspending, purifying, and preparing working aliquots of oligonucleotides (oligos) for use in molecular biology experiments. The procedure ensures the oligos are properly dissolved, purified, quantified, and aliquoted for consistent and reliable experimental results. This protocol was adapted for work conducted in the following publications: https://doi.org/10.1016/j.gene.2019.144081; https://doi.org/10.1371/journal.pone.0194205; https://doi.org/10.1016/j.arcmed.2018.09.001; https://doi.org/10.1016/j.clim.2015.12.002; https://doi.org/10.1111/iji.12355 Keywords: Oligonucleotide resuspension Oligo purification Quantitative PCR (qPCR) Molecular biology protocol DEPC water Method Overview: Resuspend lyophilized oligos in sterile DEPC H2O. Purify the oligos using sodium acetate and ethanol precipitation. Quantify the oligos with a Nanodrop and verify integrity by agarose gel electrophoresis. Aliquot the purified oligos and store at -20°C.